Catacuzzeno Luigi, Fioretti Bernard, Franciolini Fabio
Dipartimento di Biologia Cellulare e Molecolare, Università di Perugia, I-06123 Perugia, Italy.
J Neurophysiol. 2003 Dec;90(6):3688-701. doi: 10.1152/jn.00308.2003. Epub 2003 Sep 10.
A biophysical analysis of the voltage-gated K (Kv) currents of frog saccular hair cells enzymatically isolated with bacterial protease VIII was carried out, and their contribution to the cell electrical response was addressed by a modeling approach. Based on steady-state and kinetic properties of inactivation, two distinct Kv currents were found: a fast inactivating IA and a delayed rectifier IDRK. IA exhibited a strongly hyperpolarized inactivation V(1/2) (-83 mV), a relatively rapid single exponential recovery from inactivation (taurec of approximately 100 ms at -100 mV), and fast activation and deactivation kinetics. IDRK showed instead a less-hyperpolarized inactivation V(1/2) (-48 mV), a slower, double-exponential recovery from inactivation (taurec1 approximately 490 ms and taurec2 approximately 4,960 ms at -100 mV), and slower activation and deactivation kinetics. Steady-state activation gave a V(1/2) and a k of -46.2 and 8.2 mV for IA and -48.3 and 4.2 mV for IDRK. Both currents were not appreciably blocked by bath application of 10 mM TEA, but were inhibited by 4-AP, with IDRK displaying a higher sensitivity. IDRK also showed a relatively low affinity to linopirdine, being half blocked at approximately 50 microM. Steady-state and kinetic properties of IDRK and IA were described by 2nd- and 3rd-order Hodgkin-Huxley models, respectively. The goodness of our quantitative description of the Kv currents was validated by including IA and IDRK in a theoretical model of saccular hair cell electrical activity and by comparing the simulated responses with those obtained experimentally. This thorough description of the IDRK and IA will contribute toward understanding the role of these currents in the electrical response on this preparation.
对用细菌蛋白酶VIII酶解分离的青蛙球囊毛细胞的电压门控钾(Kv)电流进行了生物物理分析,并通过建模方法探讨了它们对细胞电反应的贡献。基于失活的稳态和动力学特性,发现了两种不同的Kv电流:快速失活的IA和延迟整流器IDRK。IA表现出强烈超极化的失活V(1/2)(-83 mV),从失活状态相对快速的单指数恢复(在-100 mV时taurec约为100 ms),以及快速的激活和失活动力学。相反,IDRK表现出超极化程度较小的失活V(1/2)(-48 mV),从失活状态较慢的双指数恢复(在-100 mV时taurec1约为490 ms,taurec2约为4960 ms),以及较慢的激活和失活动力学。稳态激活时,IA的V(1/2)和k分别为-46.2和8.2 mV,IDRK的为-48.3和4.2 mV。两种电流在浴液中加入10 mM四乙铵(TEA)时均未受到明显阻断,但受到4-氨基吡啶(4-AP)抑制,IDRK表现出更高的敏感性。IDRK对利诺吡啶的亲和力也相对较低,在约50 microM时被半阻断。IDRK和IA的稳态和动力学特性分别用二阶和三阶霍奇金-赫胥黎模型描述。通过将IA和IDRK纳入球囊毛细胞电活动的理论模型,并将模拟反应与实验获得的反应进行比较,验证了我们对Kv电流定量描述的准确性。对IDRK和IA的这种全面描述将有助于理解这些电流在该制剂电反应中的作用。